Expression of Selected Genes and Circulating microRNAs in Patients with Celiac Disease

Abstract

Background and Objectives: Celiac disease (CD) is an immune-mediated enteropathy with characteristic intestinal alterations. CD occurs as a chronic inflammation secondary to gluten sensitivity in genetically susceptible individuals. Until now, the exact cause of the disease has not been established, which is why new studies have appeared that address the involvement of various genes and microRNAs (miRNAs) in the pathogenesis. The aim of the study is to describe the expression of selected genes (Wnt family member 3, WNT3; Wnt family member 11, WNT11; tumor necrosis factor alpha, TNFα; mitogen-activated protein kinase 1, MAPK1; AKT serine/threonine kinase 3, AKT3; phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha, PIK3CA; and cyclin D1, CCND1) and miRNAs (miR-192-5p, miR-194-5p, miR-449a and miR-638) in adult patients with CD. Materials and Methods: In total, 15 patients with CD at diagnosis (newly diagnosed), 33 patients on a gluten-free diet (GFD) for at least 1 year and 10 controls (control) were prospectively included. Blood samples were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Results: The results show that TNFα, MAPK1 and CCND1 were significantly overexpressed (p = 0.0249, p = 0.0019 and p = 0.0275, respectively) when comparing the newly diagnosed group to the controls. The other genes studied in CD patients were mostly with high values compared to controls, without reaching statistical significance. Among the miRNAs, the closest to a statistically significant value was miR-194-5p when the newly diagnosed group versus control (p = 0.0510) and GFD group versus control (p = 0.0671) were compared. The DIANA and miRNet databases identified significant functional activity for miR-449a and miR-192-5p and an interconnection of miR-194-5p and miR-449a with CCND1. Conclusions: In conclusion, genes and circulating miRNAs require further studies as they could represent important biomarkers in clinical practice.

Keywords: biomarkers; celiac disease; circulating microRNAs; gene expression; networks.

Figure 1

Expression levels of WNT3 (a), WNT11 (b), TNFα (c), MAPK1 (d), AKT3 (e), PIK3CA (f) and CCND1 (g) in newly diagnosed CD group versus controls using qRT-PCR based on the TaqMan protocol. Abbreviations: * and ** p < 0.05.

Figure 2

Expression levels of WNT3 (a), WNT11 (b), TNFα (c), MAPK1 (d), AKT3 (e), PIK3CA (f) and CCND1 (g) in GFD CD group versus controls using qRT-PCR based on the TaqMan protocol.

Figure 3

Expression levels of WNT3 (a), WNT11 (b), TNFα (c), MAPK1 (d), AKT3 (e), PIK3CA (f) and CCND1 (g) in newly diagnosed CD versus GFD CD groups using qRT-PCR based on the TaqMan protocol.

Figure 4

Expression levels of miR-192-5p (a), miR-194-5p (b), miR-449a (c) and miR-638 (d) in newly diagnosed CD group versus controls using qRT-PCR based on the TaqMan protocol.

Figure 5

Expression levels of miR-192-5p (a), miR-194-5p (b), miR-449a (c) and miR-638 (d) in GFD CD group versus controls using qRT-PCR based on the TaqMan protocol.

Figure 6

Expression levels of miR-192-5p (a), miR-194-5p (b), miR-449a (c) and miR-638 (d) in newly diagnosed CD versus GFD groups using qRT-PCR based on the TaqMan protocol.

Figure 7

Heatmap generated using DIANA-miRPath v3.0 (http://snf-515788.vm.okeanos.grnet.gr (accessed on 5 May 2021)), showing the association with biological processes related to the evaluated miRNAs. Abbreviations: TGF, transforming growth factor; HTLV-1, human T lymphotropic virus type 1.

Figure 8

miRNA–mRNA interconnection network generated using miRNet. Abbreviations: EIF4G1, eukaryotic translation initiation factor 4 gamma 1; TP53, tumor protein p53; ARCN1, archain 1; CDC25A, cell division cycle 25A; WEE1, WEE1 G2 checkpoint kinase; PIM1, pim-1 proto-oncogene, serine/threonine kinase; ABL2, ABL proto-oncogene 2, non-receptor tyrosine kinase; KLF6, kruppel like factor 6; DDX3X, DEAD-box helicase 3 X-linked; RAB21, RAB21, member RAS oncogene family; BCL2, BCL2 apoptosis regulator; NAMPT, nicotinamide phosphoribosyltransferase; ACVR2A, activin A receptor type 2A; ALDH3B1, aldehyde dehydrogenase 3 family member B1; APP, amyloid beta precursor protein; BMI1, BMI1 proto-oncogene, polycomb ring finger.